The up-regulation of proline synthesis in the meristematic tissues of wheat seedlings upon short-term exposure to osmotic stress

An increase in the cellular concentration of free proline is a common response of many plants to various types of environmental stress. In this study, we monitored the accumulation of proline and the activities of Delta(1)-pyrroline-5-carboxylate synthetase (P5CS) and Delta(1)-pyrroline-5-carboxylate reductase (P5CR), the key enzymes of proline biosynthesis, in different parts of 4-day-old seedlings of wheat (Triticum aestivum L. cv. Josef) in the course of the first 8 h after the application of osmotic stress to determine the primary sites of proline production under water deficit conditions. Our results show that proline accumulated rapidly over this stress period in the root tips (cell division and elongation zone) and the basal region of the leaves in a time-dependent manner. Parallel to the rise in proline content, the activities of P5CS and P5CR increased markedly in these growing tissues under osmotic stress. Dissection of the root tip and the leaf base demonstrated that after 8 h of water shortage the accumulation of proline and the activities of P5CS and P5CR were highest in the regions where active cell division takes place. In the mature parts of the root and the leaf, there was virtually no enhancement of proline metabolism during the early phase of water deprivation investigated here. These data indicate that at the initial stage of water stress proline production is primarily required for the protection of the meristematic tissues in the roots and leaves. Furthermore, a transient rise in nitric oxide (NO) production was detected in the root tips and the leaf base in response to osmotic stress just before proline synthesis was enhanced. Treatment with the NO scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO) reduced considerably the increase in the activities of P5CS and P5CR and suppressed the accumulation of proline by more than 85% in the stressed root tips and the leaf base. These results suggest that NO is involved as a signalling molecule in the up-regulation of proline synthesis in the growing tissues of young wheat seedlings in response to short-term water deprivation.